G-TRACE: rapid Gal4-based cell lineage analysis in Drosophila

Evans, Cory J.; Olson, John M.; Ngo, Kathy T.; Kim, Eunha; Lee, Noemi E.; Kuoy, Edward; Patananan, Alexander N.; Sitz, Daniel; Tran, Phuong‐Thao; Do, Minh-Tu; Yackle, Kevin; Cespedes, Albert; Hartenstein, Volker; Call, Gerald B.; Banerjee, Utpal

doi:10.1038/nmeth.1356

Cited by 328 publications

(346 citation statements)

References 16 publications

(12 reference statements)

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“…We used a UAS-mCD8 GFP transgene to label the targeted cells and immunostaining against the cleaved form of Death Caspase 1 (Dcp1) to reveal cell death induction. Previous lineage-tracing experiments showed that col-GAL4 is expressed solely in the PSC in the primary lobes and sporadically in posterior lobes (17), whereas Antp-GAL4 is expressed in the PSC as well as in some cardiomyocytes present between the primary lobes (20) (Fig. 1 and Figs.…”

Section: Resultsmentioning

confidence: 87%

The EBF transcription factor Collier directly promotes Drosophila blood cell progenitor maintenance independently of the niche

Benmimoun

Polesello

Haenlin

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

149

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The maintenance of stem or progenitor cell fate relies on intrinsic factors as well as local cues from the cellular microenvironment and systemic signaling. In the lymph gland, an hematopoietic organ in Drosophila larva, a group of cells called the Posterior Signaling Centre (PSC), whose specification depends on the EBF transcription factor Collier (Col) and the HOX factor Antennapedia (Antp), has been proposed to form a niche required to maintain the pool of hematopoietic progenitors (prohemocytes). In contrast with this model, we show here that genetic ablation of the PSC does not cause an increase in blood cell differentiation or a loss of blood cell progenitors. Furthermore, although both col and Antp mutant larvae are devoid of PSC, the massive prohemocyte differentiation observed in col mutant is not phenocopied in Antp mutant. Interestingly, beside its expression in the PSC, Col is also expressed at low levels in prohemocytes and we show that this expression persists in PSC-ablated and Antp mutant larvae. Moreover, targeted knockdown and rescue experiments indicate that Col expression is required in the prohemocytes to prevent their differentiation. Together, our findings show that the PSC is dispensable for blood cell progenitor maintenance and reveal the key role of the conserved transcription factor Col as an intrinsic regulator of hematopoietic progenitor fate.hematopoiesis | Drosophila | stem cell niche | EBF

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Section: Resultsmentioning

confidence: 87%

The EBF transcription factor Collier directly promotes Drosophila blood cell progenitor maintenance independently of the niche

Benmimoun

Polesello

Haenlin

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

149

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“…Thus, our system can also be used as a lineage-tracing tool reminiscent of the previously reported G-trace system (29). Importantly, a tubGal80ts allele can be used to repress early Gal4 activity and achieve "real-time" capture of cell-cell interactions (Fig.…”

Section: Resultsmentioning

confidence: 96%

Development of an optimized synthetic Notch receptor as an in vivo cell–cell contact sensor

Huang

Perrimon

2017

Proc. Natl. Acad. Sci. U.S.A.

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Detection and manipulation of direct cell-cell contact in complex tissues is a fundamental and challenging problem in many biological studies. Here, we report an optimized Notch-based synthetic receptor (synNQ) useful to study direct cell-cell interactions in Drosophila. With the synNQ system, cells expressing a synthetic receptor, which contains Notch activation machinery and a downstream transcriptional activator, QF, are activated by a synthetic GFP ligand expressed by contacting neighbor cells. To avoid cis-inhibition, mutually exclusive expression of the synthetic ligand and receptor is achieved using the "flippase-out" system. Expression of the synthetic GFP ligand is controlled by the Gal4/UAS system for easy and broad applications. Using synNQ, we successfully visualized cell-cell interactions within and between most fly tissues, revealing previously undocumented cell-cell contacts. Importantly, in addition to detection of cells in contact with one another, synNQ allows for genetic manipulation in all cells in contact with a targeted cell population, which we demonstrate in the context of cell competition in developing wing disks. Altogether, the synNQ genetic system will enable a broad range of studies of cell contact in developmental biology.

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“…Flies carrying an enhancerless GFP construct downstream of an hsp70 minimal promoter at recombinase-mediated cassette exchange (RMCE) site 53F were described pre- Evans et al 2009), carrying an FRT-flanked stop cassette between a ubiquitous promoter and a nuclear EGFP coding region on chromosome 3 ("G-TRACE cassette"), were obtained from the Bloomington Drosophila Stock Center (stock no. 32251).…”

Section: Stocks and Fly Husbandrymentioning

confidence: 99%

“…We therefore wished to provide further confirmation that these enhancers indeed activate promoters in trans. To this end, we designed a complementary system to detect transvection based on the G-TRACE tools for cell lineage analysis (Evans et al 2009). In this system, the recombinase FLP acts on a G-TRACE cassette to remove a transcriptional stop signal between a ubiquitous enhancer and a GFP transgene, resulting in robust GFP fluorescence in a cell's lineage ( Figure 3A).…”

Section: A Flp-based Assay Further Supports Transvection By Weak Clasmentioning

confidence: 99%

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The Capacity to Act in Trans Varies Among Drosophila Enhancers

Blick

Mayer-Hirshfeld

Malibiran³

et al. 2016

Genetics

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The interphase nucleus is organized such that genomic segments interact in cis, on the same chromosome, and in trans, between different chromosomes. In Drosophila and other Dipterans, extensive interactions are observed between homologous chromosomes, which can permit enhancers and promoters to communicate in trans. Enhancer action in trans has been observed for a handful of genes in Drosophila, but it is as yet unclear whether this is a general property of all enhancers or specific to a few. Here, we test a collection of well-characterized enhancers for the capacity to act in trans. Specifically, we tested 18 enhancers that are active in either the eye or wing disc of third instar Drosophila larvae and, using two different assays, found evidence that each enhancer can act in trans. However, the degree to which trans-action was supported varied greatly between enhancers. Quantitative analysis of enhancer activity supports a model wherein an enhancer's strength of transcriptional activation is a major determinant of its ability to act in trans, but that additional factors may also contribute to an enhancer's trans-activity. In sum, our data suggest that a capacity to activate a promoter on a paired chromosome is common among Drosophila enhancers.KEYWORDS RMCE; interchromosomal interactions; long-range enhancer; somatic homolog pairing; transvection T HE spatial organization of the interphase eukaryotic genome is characterized by extensive long-distance interactions between distal chromosome regions (Sanyal et al. 2012). Interactions have been identified between sequences on the same chromosome (in cis) or on different chromosomes (in trans) (Lieberman-Aiden et al. 2009;Duan et al. 2010;Sexton et al. 2012;van de Werken et al. 2012;Nagano et al. 2013;Zhang et al. 2013). Many long-distance interactions in cis underlie the activation of specific genes, and in some cases, sequences have been identified that facilitate interactions between a distal enhancer and a specific promoter target (Zhou and Levine 1999;Calhoun et al. 2002;Calhoun and Levine 2003;Lin 2003;Akbari et al. 2008;Fujioka et al. 2009;Majumder et al. 2015). In contrast, the genetic impacts of trans-interactions between chromosomes are less clearly understood. Examples of gene regulation involving interchromosomal associations have been described (Spilianakis et al. 2005;Bacher et al. 2006;Xu et al. 2006;Apostolou and Thanos 2008;Sandhu et al. 2009;Markenscoff-Papadimitriou et al. 2014;Patel et al. 2014), but it remains unclear whether it is common for sequences that regulate gene expression to communicate between different chromosomes when they are physically juxtaposed.In Drosophila melanogaster, extensive trans-interactions are observed between homologous chromosomes in virtually all somatic tissues, a phenomenon known as somatic homolog pairing (reviewed by McKee 2004;Bosco 2012). The close proximity of homologous chromosomes in Drosophila can permit an enhancer to act in trans on a promoter on the paired homolog, a form of pairing-dependent ...

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G-TRACE: rapid Gal4-based cell lineage analysis in Drosophila

Cited by 328 publications

References 16 publications

The EBF transcription factor Collier directly promotes Drosophila blood cell progenitor maintenance independently of the niche

The EBF transcription factor Collier directly promotes Drosophila blood cell progenitor maintenance independently of the niche

Development of an optimized synthetic Notch receptor as an in vivo cell–cell contact sensor

The Capacity to Act in Trans Varies Among Drosophila Enhancers

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